• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.04a
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• pp.44-44
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• 2019

This study aimed to develop a suitable method for inducing the proliferation of prothallus and producing sporophytes of rock polypody (Polypodium vulgare L.). The prothalli used in all experiments were obtained from spore germination and sub-cultured for 8-week intervals. The most appropriate media for prothallus propagation were investigated by culturing 300 mg of prothallus in MS ($1/4{\times}$, $1/2{\times}$, $1{\times}$, and $2{\times}$ strength) medium and in Knop medium for 8 weeks. Cultures were maintained at a temperature of $25{\pm}1^{\circ}C$, light intensity of $30{\pm}1.0{\mu}mol-m-2{\cdot}s-1$, and a photoperiod of 16/8 h (light/dark). Fresh weight of prothalli was 4.8 g on $1{\times}$ MS, 4.5 g on $1/2{\times}$ MS and 4.3 g on 1/4 MS medium. To select a suitable soil combination for sporophyte formation, 1.0 g of prothallus was ground with distilled water, spread in five combinations onto different soil substrates (decomposed granite, horticultural substrates, peat moss, and perlite), and then cultivated for 13 weeks. The sporophyte cultures were maintained at a temperature of $25{\pm}1^{\circ}C$, light intensity of $43{\pm}2.0{\mu}mol-m-2{\cdot}s-1$, humidity of $84{\pm}1.4%$, and a photoperiod of 16/8 h (light/dark). The results showed that a mixture containing a 2:1 (v:v) ratio of horticultural substrate and perlite, increased sporophyte formation to 462.5 sporophytes per pot (7.5 cm2). The other soil substrates produced from 314.5 to 405.3 sporophytes per pot. Therefore, our results will provide conditions suitable for mass production of Polypodium vulgare L.

• Journal of Microbiology and Biotechnology
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• v.32 no.2
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• pp.160-169
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• 2022

In the present study we evaluated the efficacy of a bioformulation of Streptomyces cameroonensis for control of black pod disease in cocoa and enhancement of seedling growth. The formulation developed using talc powder and cassava starch as carriers showed high shelf-life of 1.07 × 10⁶ CFU/g after six months storage at 4℃. The formulation was tested for inhibition of spore germination in Phytophthora megakarya and showed 100% inhibition at 10% (w/v) of formulation. To determine the efficacy of the formulation, we performed an in planta assay in the greenhouse on two hybrids of cocoa seedlings, the tolerant SNK413 × (♂) T79/467 and the susceptible UPA 134× (♂) SCA 12. Detached leaf assay showed a significant reduction in the disease severity index of about 67% for the tolerant hybrid and 55% for the susceptible hybrid compared to non-treated plants. A significant enhancement in stem length, leaf surface area and root weight was observed. Analysis of biochemical markers of defense showed a significant increase in total polyphenol, flavonoid, and total protein contents. There was also significant upregulation of PR-proteins such as chitinases, peroxidases and β-1, 3-glucanases following treatment of both tolerant and susceptible hybrids, though with a higher level of synthesis in the tolerant hybrids. A significant increase was also observed in polyphenol oxidase activities in plants treated with the formulation. This work demonstrated the stability and effectiveness of the S. cameroonensis powder formulation in suppressing black pod disease in cocoa and subsequently enhancing the growth of seedlings.

• The Plant Pathology Journal
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• v.37 no.6
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• pp.533-542
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• 2021

Chitosan has been considered an environmental-friendly polymer. However, its use in agriculture has not been extended yet due to its relatively low solubility in water. N-Methylene phosphonic chitosan (NMPC) is a water-soluble derivative prepared by adding a phosphonic group to chitosan. This study demonstrates that NMPC has a fungicidal effect on the phytopathogenic fungus Fusarium solani f. sp. eumartii (F. eumartii) judged by the inhibition of F. eumartti mycelial growth and spore germination. NMPC affected fungal membrane permeability, reactive oxygen species production, and cell death. Also, this chitosan-derivative exerted antifungal effects against two other phytopathogens, Botrytis cinerea, and Phytophthora infestans. NMPC did not affect tomato cell viability at the same doses applied to these phytopathogens to exert fungicide action. In addition to water solubility, the selective biological cytotoxicity of NMPC adds value in its application as an antimicrobial agent in agriculture.

• Journal of Microbiology and Biotechnology
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• v.33 no.5
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• pp.607-620
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• 2023

The biocontrol approach using beneficial microorganisms to control crop diseases is becoming an essential alternative to chemical fungicides. Therefore, new and efficient biocontrol agents (BCA) are needed. In this study, a rhizospheric actinomycete isolate showed unique and promising antagonistic activity against three of the most common phytopathogenic fungi, Fusarium oxysporum MH105, Rhizoctonia solani To18, and Alternaria brassicicola CBS107. Identification of the antagonistic strain, which was performed according to spore morphology and cell wall chemotype, suggested that it belongs to the Nocardiopsaceae. Furthermore, cultural, physiological, and biochemical characteristics, together with phylogenetic analysis of the 16S rRNA gene (OP869859.1), indicated the identity of this strain to Nocardiopsis alba. The cell-free filtrate (CFF) of the strain was evaluated for its antifungal potency, and the resultant inhibition zone diameters ranged from 17.0 ± 0.92 to 19.5 ± 0.28 mm for the tested fungal species. Additionally, the CFF was evaluated in vitro to control Fusarium wilt disease in Vicia faba using the spraying method under greenhouse conditions, and the results showed marked differences in virulence between the control and treatment plants, indicating the biocontrol efficacy of this actinomycete. A promising plant-growth promoting (PGP) ability in seed germination and seedling growth of V. faba was also recorded in vitro for the CFF, which displayed PGP traits of phosphate solubilization (48 mg/100 ml) as well as production of indole acetic acid (34 ㎍/ml) and ammonia (20 ㎍/ml). This study provided scientific validation that the new rhizobacterium Nocardiopsis alba strain BH35 could be further utilized in bioformulation and possesses biocontrol and plant growth-promoting capabilities.

• Journal of Microbiology and Biotechnology
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• v.34 no.5
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• pp.1040-1050
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• 2024

To isolate and analyze bacteria with Verticillium wilt-resistant properties from the fermentation residue of kitchen wastes, as well as explore their potential for new applications of the residue. A total of six bacterial strains exhibiting Verticillium wilt-resistant capabilities were isolated from the biogas residue of kitchen waste fermentation. Using a polyphasic approach, strain ZL6, which displayed the highest antagonistic activity against cotton Verticillium wilt, was identified as belonging to the Pseudomonas aeruginosa. Bioassay results demonstrated that this strain possessed robust antagonistic abilities, effectively inhibiting V. dahliae spore germination and mycelial growth. Furthermore, P. aeruginosa ZL6 exhibited high temperature resistance (42℃), nitrogen fixation, and phosphorus removal activities. Pot experiments revealed that P. aeruginosa ZL6 fermentation broth treatment achieved a 47.72% biological control effect compared to the control group. Through activity tracking and protein mass spectrometry identification, a neutral metalloproteinase (Nml) was hypothesized as the main virulence factor. The mutant strain ZL6ߡNml exhibited a significant reduction in its ability to inhibit cotton Verticillium wilt compared to the strain P. aeruginosa ZL6. While the inhibitory activities could be partially restored by a complementation of nml gene in the mutant strain ZL6CMߡNml. This research provides a theoretical foundation for the future development and application of biogas residue as biocontrol agents against Verticillium wilt and as biological preservatives for agricultural products. Additionally, this study presents a novel approach for mitigating the substantial amount of biogas residue generated from kitchen waste fermentation.

• Research in Plant Disease
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• v.18 no.3
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• pp.201-209
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• 2012

The multi-function of 18 Bacillus subtilis isolates collected from agricultural extension centers of local government and National Academy of Agricultural Science was investigated by measuring their antifungal activities against five plant pathogens, such as Rhizoctonia solani, Colletotrichum acutatum, Fusarium oxysporum, Magnaporthe oryzae and Phytophthora capsici, phosphorus solubilization ability, production of indole acetic acid (IAA) and siderophore, and nitrogen fixation. The B. subtilis isolates showed antifungal activity against several plant pathogens and nitrogen fixation activity, and produced siderophore and IAA. They could control pepper powdery mildew (Leveillula taurica), but there was no difference in control efficacy among the B. subtilis isolates. In fields, the control efficacy of B. subtilis R2-1 ($10^8$ cells/ml) was compared with two microbial fungicides, Q-pect and Topsid. In 2009, the control efficacy of B. subtilis R2-1 (37.7%) was lower than that of Topsid (47.6%), but higher than that of Q-pect (25.7%). In 2010, the control efficacy of B. subtilis R2-1 (83.3%) was higher than that of Topsid (67.9%). In order to elucidate mode of action of B. subtilis R2-1 for controlling pepper powdery mildew, spore germination rates of pepper powdery mildew pathogen collected on treated leaves was investigated when suspensions of B. subtilis R2-1 and two microbial fungicides (Q-pect and Topsid) were foliar-sprayed. They highly suppressed spore germination of the pathogen with inhibition values of 84.2% for B. subtilis R2-1, 97.9% for Q-pect and 94.7% for Topsid. Further study on the mass-culturing method and formulation is needed for development of a microbial fungicide.

• The Korean Journal of Pesticide Science
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• v.18 no.2
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• pp.115-121
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• 2014

This study was performed to investigate the effect of conidial density, wetness period and temperature on conidial germination, appressoria formation and disease incidence. While there was not significantly correlated between conidial density and temperature, and conidial germination and appressoria formation, there was a significant correlation between those factors and disease incidence. The longer wetness period was, the higher the ratio of conidial germination, appressoria formation and the disease incidence was. The optimum conidial density, temperature and wetness period was $1{\times}10^6$ conidia $mL^{-1}$, $30^{\circ}C$ and 5 days, respectively. In case the wetness period was more than 5 days, the typical symptom was not found on pepper fruits because of the overgrowth of mycelia. Using this fruit assay method, which the pepper anthracnose pathogens were inoculated by spraying spore suspension on non-wounded or wounded pepper fruits, control effect of three fungicides were evaluated against pepper anthracnose by the protective and/or the curative application. Propineb showed high protective control activity, while it showed curative control activity on unwounded fruits, but did not showed curative control activity on wounded fruits. Tebuconazole, one of curative fungicide, showed higher control activity in non-wound inoculation than wound inoculation. Trifloxystrobin, one of strobilurin group, showed high both protective and control activity against anthracnose. In conclusion, we supposed that the newly developed in vitro pepper fruit assay can be used to evaluate antifungal activity of control agents against pepper anthracnose.

• The Korean Journal of Pesticide Science
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• v.5 no.2
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• pp.37-44
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• 2001

The biological effects of the iminoctadine tris (albesilate) and kresoxim-methyl for the protection of citrus postharvest diseases caused by penicillium spp. were assayed. In vitro tests, $EC_{50}$ values of iminoctadine tris(albesilate) were $0.01{\sim}0.02\;and\;0.01{\mu}g$ a.i./mL against mycelial growth of P. italicum and P. digitatum, respectively, but iminoctadine tris(albesilate) at $0.64{\mu}g$ a.i. /mL inhibited a little mycelial growth of unknown Penicillium sp. which produced another symptom different to blue and green mold caused by P. italicum and P. digitatum, respectively. And against germination and growth of germ tube of P. italicum and P. digitatum, $EC_{50}$ value of iminoctadine tris(albesilate) was $0.0013{\sim}0.0025{\mu}g$ a.i./mL. But spore germination of unknown Penicillium spp. was not nearly inhibited at $0.2{\mu}g$ a.i./mL. $EC_{50}$ values of kresoxim-methyl were $0.08{\sim}0.16$, 0.04 and $0.16{\mu}g$ a.i./mL against mycelial growth of P. italicum, P. digitatum and unknown Penicillium sp., respectively, and $0.04{\sim}0.08{\mu}g$ a.i./mL and $0.01{\sim}0.02{\mu}g$ a.i./mL against germination and growth of germ tube of P. italicum and unknown Penicillium sp., and P. digitatum, respectively. Iminoctadine tris(albesilate) and kresoxim-methyl were markedly effective to control the postharvest disease by 7 days spray prior to harvest. When the fruits were sprayed with iminoctadine-tris(albesilate) ($200{\mu}g$ a.i./mL) and kresoxim-methyl ($155{\mu}g$ a.i./mL) 7 days prior to harvest and subsequently stored for 90 days, the percentage of diseased fruit by Penicillium spp. was $3.6{\pm}1.8%$ in treatment of kresoxim-methyl and $5.9{\pm}1.8%$ in iminoctadine-tris(albesilate), respectively. On the other hand, tile percentage of diseased fruit was relatively high, $20.3{\pm}10.0%\;and\;19.5{\pm}9.6%$ in thiophanate-methyl ($700{\mu}g$ a.i./mL) and non-treatment, respectively. Maximum residue amount (ppm) among fruits (flesh and peel) assayed 0, 30, 60 and 90 days after storage was 0.45 and 0.10 ppm in treatment of kresoxim-methyl and iminoctadine, respectively.

• Microbiology and Biotechnology Letters
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• v.37 no.3
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• pp.280-286
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• 2009

To investigate anti-phytopathogenic fungal activity of Coptis chinensis, the methanol extract and its organic solvent fractions were prepared. Using the extract and the fractions, in-vitro spore-germination inhibition and mycelial-growth inhibition activities were evaluated against Colletotrichum gloeosporioides, Phytohpthora capsici, Pyricularia grisea, Rhizoctonia solani, Botryosphaeri dothidea, Glomerella cingulata, respectively. Treatment of the methanol extract (500 mg/mL) into the spore of phytopathogenic fungi completely inhibited germinations for 5 days, except B. dothidea, and showed strong antifungal activities against P. grisea and B. cinerea, and antioomycetes activity against P. capsici. The minimal growth inhibition concentrations of the methanol extract against P. grisea, B. cinerea and P. capsici were 300, 300, and 500 mg/mL, respectively. For practical application of C. chinensis in red-pepper field, the hot-water extract (1,000 mg/mL) was prepared in commercial facility, after evaluation of heat stability and solvent-extraction yields of antifungal substances. The 3-times leaf-spray of the extract from June to August, 2008 did not show any deleterious effect to red-pepper. In fact, the leaf-spray promoted plant growth including leaf, root and fruit. The average weight and rind of each fruit were increased to 119% and 117% comparison to those of without treatments. Our results suggest that C. chinensis is a useful source for control of red-pepper diseases and plant growth.

• Korean Journal of Agricultural Science
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• v.13 no.1
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• pp.52-61
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• 1986

Mycelial growth of Colletotrichun dematium and C. gloeosporioides isolated from anthracnose of Lycium chinense was favorable on V-8 Juice Agar media and Oatmeal Agar media of the four different media tested. Sporulation of C. dematium was favorable on PDA media and V-8 Juice Agar media and that of C. gloeosporioides was favorable on all media tested. The optimum temperature for mycelial growth of C. gloeosporioides and C. dematium was $25^{\circ}C$ and $28^{\circ}C$, respectively and sporulation of the fungi was stimulated under alternating NUV light and darkness. Of the seven nitrogen sources tested, gelatin was the most favorable for mycelial growth of C. gloeosporioides and glycin and gelatin were favorable for that of C. dematium. Of the eleven carbon sources tested, dextrin and sorbitol were favorable for mycelial growth of C. gloeosporioides and dextrin was the most favorable for that of C. dematium. Among the eleven fugicides tested, Benomyl, Folpet, Dithianon, Carbendazim and S-3308L were found superior for the inhibition of mycelial growth and spore germination of the fungi. They were, also, found superior for the control of anthracnose disease of L. chinense when they were applied to the plants in the field.